Difference between revisions of "Part:BBa K255001"
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===Result===
 
===Result===
−
The YFP Expression increased around 10 folds from 0 to 100 μM of IPTG Conc and thereafter its value was almost constant to 200 and 500 μM of IPTG conc.
+
The YFP Expression increased around 10 folds from 0 to 100 μM of IPTG Conc and thereafter its value was almost constant to 200 and 500 μM of IPTG conc (fig.1).
−
[[Image:p1.png|thumb|600px|center|alt=A |Expression of BBa_K255001 in lacI deleted E.coli strain]]
+
[[Image:p1.png|thumb|600px|center|alt=A |Figure 1. Expression of BBa_K255001 in lacI deleted E.coli strain]]
  
−
After this the expression profile of YFP between 0 and 100 μM was measured.
+
After this the expression profile of YFP between 0 and 100 μM was measured(fig.2).
 +
 
 +
[[Image:p1_2.png|thumb|200px|center|alt=A |Figure 2. Expression of BBa_K255001 in lacI deleted E.coli strain]]
 +
As a control, FACS was done with host strain to quantitate  the autofluorescence. It was observed that cells are showing autofluorescence  but its  intensity is very less than our BBa_K255001 strain (fig.3)
 +
[[Image:auto.png|thumb|200px|center|alt=A |Figure 3. Autofluorescence]]
  
−
[[Image:p1_2.png|thumb|200px|center|alt=A |Expression of BBa_K255001 in lacI deleted E.coli strain]]
 
 
===Safety===
 
===Safety===
−
Because this part is from commonly used elements in bacteria. No safety problem can be arised by this part.  
+
Because this part is from commonly used elements in bacteria. No safety problem can be arisen by this part.  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 19:31, 31 October 2009

pLac.lacI-cfp(fusion).terminator.ptet.yfp.teminator

It has dual negative feedback loop one on the plasmid copy number and second on the LacI expression. This network is present in IPTG inducible plasmid,pSB2K3. Both LacI and plasmid copy number are dependent on IPTG concentration (inducer). The copy number is characterized by YFP while LacI is characterized by LacI-CFP fusion protein.

Characterization

YFP was measured using FACS analysis. Illustrative FACS results are given below: The mean, variance and standard error were noted for difference strains and at different IPTG concentration. The FACS results are shown for varying IPTG concentrations varying from 0um to 500um.

Result

The YFP Expression increased around 10 folds from 0 to 100 μM of IPTG Conc and thereafter its value was almost constant to 200 and 500 μM of IPTG conc (fig.1).

A
Figure 1. Expression of BBa_K255001 in lacI deleted E.coli strain

After this the expression profile of YFP between 0 and 100 μM was measured(fig.2).

A
Figure 2. Expression of BBa_K255001 in lacI deleted E.coli strain
As a control, FACS was done with host strain to quantitate  the autofluorescence. It was observed that cells are showing autofluorescence  but its  intensity is very less than our BBa_K255001 strain (fig.3)
A
Figure 3. Autofluorescence

Safety

Because this part is from commonly used elements in bacteria. No safety problem can be arisen by this part.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1162
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]